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流浪转座元件在果蝇物种中具有转座酶依赖性和非依赖性切除活性。

The hobo transposable element has transposase-dependent and -independent excision activity in drosophilid species.

作者信息

Handler A M, Gomez S P

机构信息

Insect Attractants, Behavior and Basic Biology Research Laboratory, U.S. Department of Agriculture, Gainesville, Florida 32608, USA.

出版信息

Mol Gen Genet. 1995 May 20;247(4):399-408. doi: 10.1007/BF00293140.

Abstract

Mobility of the hobo transposable element was determined for several strains of Drosophila melanogaster and several Drosophila species. Mobility was assessed by use of an in vivo transient assay in the soma of developing embryos, which monitored hobo excision from injected indicator plasmids. Excision was detected in a D. melanogaster strain (cn; ry42) devoid of endogenous hobo elements only after co-injection of a helper plasmid containing functional hobo transposase under either heat shock or normal promoter regulation. Excision was also detected in D. melanogaster without helper in strains known to contain genomic copies of hobo. In Drosophila species confirmed not to contain hobo, hobo excision occurred at significant rates both in the presence and absence of co-injected helper plasmid. In four of the seven species tested, excision frequencies were two- to fivefold lower in the presence of plasmid-borne hobo, hobo excision donor sites were sequenced in indicator plasmids extracted from D. melanogaster cn; ry42 and D. virilis embryos. In the presence of hobo transposase, the predominant excision sites were identical in both species, having breakpoints at the hobo termini with an inverted duplication of proximal insertion site DNA. However, in the absence of hobo transposase in D. virilis, excision breakpoints were apparently random and occurred distal to the hobo termini. The data indicate that hobo is capable of functioning in the soma during embryogenesis, and that its mobility is unrestricted in drosophilids. Furthermore, drosophilids not containing hobo are able to mobilize hobo, presumably by a hobo-related cross-mobilizing system. The cross-mobilizing system in D. virilis is not functionally identical to hobo with respect to excision sequence specificity.

摘要

在几种黑腹果蝇品系和几个果蝇物种中测定了hobo转座元件的移动性。通过在发育中胚胎的体细胞中进行体内瞬时测定来评估移动性,该测定监测从注射的指示质粒中切除hobo的情况。仅在热休克或正常启动子调控下共注射含有功能性hobo转座酶的辅助质粒后,才在不含内源性hobo元件的黑腹果蝇品系(cn; ry42)中检测到切除。在已知含有hobo基因组拷贝的黑腹果蝇品系中,无辅助质粒时也检测到了切除。在确认不含hobo的果蝇物种中,无论是否共注射辅助质粒,hobo切除都以显著速率发生。在测试的七个物种中的四个中,存在质粒携带的hobo时切除频率降低了两到五倍,从黑腹果蝇cn; ry42和粗壮果蝇胚胎中提取的指示质粒中的hobo切除供体位点进行了测序。在存在hobo转座酶的情况下,两个物种中的主要切除位点相同,在hobo末端有断点,近端插入位点DNA有反向重复。然而,在粗壮果蝇中不存在hobo转座酶时,切除断点显然是随机的,且发生在hobo末端的远端。数据表明,hobo在胚胎发生期间能够在体细胞中发挥作用,并且其移动性在果蝇中不受限制。此外,不含hobo的果蝇能够动员hobo,推测是通过与hobo相关的交叉动员系统。就切除序列特异性而言,粗壮果蝇中的交叉动员系统在功能上与hobo不同。

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